Redesign of the coenzyme specificity of a dehydrogenase by protein engineering.
Directed mutagenesis and molecular modelling have been used to identify a set of amino-acid side chains in glutathione reductase that confer specificity for the coenzyme NADP+. Systematic replacement of these amino acids, all of which occur in a 'fingerprint' structural motif in the NADP+-binding domain, leaves the substrate specificity unchanged but converts the enzyme into one displaying a marked preference for the coenzyme NAD+.
Scrutton
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NS
Department of Biochemistry, University of Cambridge, UK.
Berry
A
A
Perham
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RN
D000595:Amino Acid Sequence; D001665:Binding Sites; D003196:Computer Graphics; D004252:DNA Mutational Analysis; D005980:Glutathione Reductase / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism*; D009249:NADP / Q000378:metabolism*; D015202:Protein Engineering; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D009243:NAD; D009249:NADP; D005980:Glutathione Reductase